NOS3 Antibody | Affinity Biosciences

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Product:	NOS3 Antibody
Catalog:	DF6450
Description:	Rabbit polyclonal antibody to NOS3
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Rabbit, Dog
Mol.Wt.:	133kDa; 133kD(Calculated).
Uniprot:	P29474
RRID:	AB_2838413

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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Related Downloads

MS Report

HPLC Report

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Prediction:

Pig(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(88%)

Clonality:

Polyclonal

Specificity:

NOS3 Antibody detects endogenous levels of total NOS3.

RRID:

AB_2838413
Cite Format: Affinity Biosciences Cat# DF6450, RRID:AB_2838413.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

cNOS; Constitutive NOS; EC NOS; EC-NOS; ecNOS; Endothelial nitric oxidase synthase; Endothelial nitric oxide synthase; Endothelial nitric oxide synthase 3; Endothelial NOS; eNOS; Nitric oxide synthase 3 (endothelial cell); Nitric oxide synthase 3; Nitric oxide synthase 3 endothelial cell; Nitric oxide synthase endothelial; Nitric oxide synthase, endothelial; NOS 3; NOS III; NOS type III; NOS3; NOS3_HUMAN; NOSIII;

Immunogens

Immunogen:

Uniprot:

P29474(NOS3_HUMAN) >>Visit HPA database.

Gene(ID):

NOS3(4846)

Expression:

P29474 NOS3_HUMAN:

Platelets, placenta, liver and kidney.

Description:

Endothelial nitric-oxide synthase (eNOS) is an important enzyme in the cardiovascular system. It catalyzes the production of nitric oxide (NO), a key regulator of blood pressure, vascular remodeling, and angiogenesis (1,2). The activity of eNOS is regulated by phosphorylation at multiple sites. The two most thoroughly studied sites are the activation site Ser1177 and the inhibitory site Thr495 (3). Several protein kinases including Akt/PKB, PKA, and AMPK activate eNOS by phosphorylating Ser1177 in response to various stimuli (4,5). In contrast, bradykinin and H2O2 activate eNOS activity by promoting both Ser1177 phosphorylation and Thr495 dephosphorylation (6,7). eNOS is activated by VEGF, and this activation is associated with dephsophorylation of eNOS at serine 113. Cyclosporin A blocks this dephosphorylation of eNOS upon VEGF stimulation (8).

Sequence:

P29474 NOS3_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MGNLKSVAQEPGPPCGLGLGLGLGLCGKQGPATPAPEPSRAPASLLPPAPEHSPPSSPLTQPPEGPKFPRVKNWEVGSITYDTLSAQAQQDGPCTPRRCLGSLVFPRKLQGRPSPGPPAPEQLLSQARDFINQYYSSIKRSGSQAHEQRLQEVEAEVAATGTYQLRESELVFGAKQAWRNAPRCVGRIQWGKLQVFDARDCRSAQEMFTYICNHIKYATNRGNLRSAITVFPQRCPGRGDFRIWNSQLVRYAGYRQQDGSVRGDPANVEITELCIQHGWTPGNGRFDVLPLLLQAPDDPPELFLLPPELVLEVPLEHPTLEWFAALGLRWYALPAVSNMLLEIGGLEFPAAPFSGWYMSTEIGTRNLCDPHRYNILEDVAVCMDLDTRTTSSLWKDKAAVEINVAVLHSYQLAKVTIVDHHAATASFMKHLENEQKARGGCPADWAWIVPPISGSLTPVFHQEMVNYFLSPAFRYQPDPWKGSAAKGTGITRKKTFKEVANAVKISASLMGTVMAKRVKATILYGSETGRAQSYAQQLGRLFRKAFDPRVLCMDEYDVVSLEHETLVLVVTSTFGNGDPPENGESFAAALMEMSGPYNSSPRPEQHKSYKIRFNSISCSDPLVSSWRRKRKESSNTDSAGALGTLRFCVFGLGSRAYPHFCAFARAVDTRLEELGGERLLQLGQGDELCGQEEAFRGWAQAAFQAACETFCVGEDAKAAARDIFSPKRSWKRQRYRLSAQAEGLQLLPGLIHVHRRKMFQATIRSVENLQSSKSTRATILVRLDTGGQEGLQYQPGDHIGVCPPNRPGLVEALLSRVEDPPAPTEPVAVEQLEKGSPGGPPPGWVRDPRLPPCTLRQALTFFLDITSPPSPQLLRLLSTLAEEPREQQELEALSQDPRRYEEWKWFRCPTLLEVLEQFPSVALPAPLLLTQLPLLQPRYYSVSSAPSTHPGEIHLTVAVLAYRTQDGLGPLHYGVCSTWLSQLKPGDPVPCFIRGAPSFRLPPDPSLPCILVGPGTGIAPFRGFWQERLHDIESKGLQPTPMTLVFGCRCSQLDHLYRDEVQNAQQRGVFGRVLTAFSREPDNPKTYVQDILRTELAAEVHRVLCLERGHMFVCGDVTMATNVLQTVQRILATEGDMELDEAGDVIGVLRDQQRYHEDIFGLTLRTQEVTSRIRTQSFSLQERQLRGAVPWAFDPPGSDTNSP

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Horse

100

Bovine

100

Rabbit

100

Dog

Sheep

Xenopus

Zebrafish

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P29474 As Substrate

P29474

Site	PTM Type	Enzyme	Source
K5	Methylation		Uniprot
T33	Phosphorylation		Uniprot
S53	Phosphorylation		Uniprot
Y81	Phosphorylation		Uniprot
C94	S-Nitrosylation		Uniprot
C99	S-Nitrosylation		Uniprot
S102	Phosphorylation		Uniprot
S114	Phosphorylation	Q00535 (CDK5)	Uniprot
C184	S-Nitrosylation		Uniprot
C201	S-Nitrosylation		Uniprot
Y210	Phosphorylation		Uniprot
C212	S-Nitrosylation		Uniprot
S260	Phosphorylation		Uniprot
T495	Phosphorylation	Q04759 (PRKCQ) , Q13237 (PRKG2) , P05771 (PRKCB) , P17612 (PRKACA) , Q02156 (PRKCE) , O75116 (ROCK2) , P17252 (PRKCA) , P05129 (PRKCG) , P24723 (PRKCH) , Q05655 (PRKCD) , Q13131 (PRKAA1) , P41743 (PRKCI) , Q05513 (PRKCZ)	Uniprot
S508	Phosphorylation		Uniprot
T512	Phosphorylation		Uniprot
S615	Phosphorylation	P17612 (PRKACA) , Q9Y243 (AKT3) , P31751 (AKT2) , P31749 (AKT1)	Uniprot
S633	Phosphorylation	Q13237 (PRKG2) , P54646 (PRKAA2) , Q13131 (PRKAA1) , P17612 (PRKACA)	Uniprot
S634	Phosphorylation		Uniprot
T636	Phosphorylation		Uniprot
S638	Phosphorylation		Uniprot
Y657	Phosphorylation	Q14289 (PTK2B)	Uniprot
C661	S-Nitrosylation		Uniprot
S738	Phosphorylation		Uniprot
K773	Ubiquitination		Uniprot
C802	S-Nitrosylation		Uniprot
K834	Ubiquitination		Uniprot
S836	Phosphorylation		Uniprot
C853	S-Nitrosylation		Uniprot
T854	Phosphorylation		Uniprot
S870	Phosphorylation		Uniprot
K904	Ubiquitination		Uniprot
C976	S-Nitrosylation		Uniprot
C991	S-Nitrosylation		Uniprot
K1035	Ubiquitination		Uniprot
C1048	S-Nitrosylation		Uniprot
C1050	S-Nitrosylation		Uniprot
K1085	Ubiquitination		Uniprot
T1094	Phosphorylation		Uniprot
C1114	S-Nitrosylation		Uniprot
T1175	Phosphorylation		Uniprot
S1177	O-Glycosylation		Uniprot
S1177	Phosphorylation	P22612 (PRKACG) , Q9Y478 (PRKAB1) , P31749 (AKT1) , Q13131 (PRKAA1) , P54646 (PRKAA2) , Q13237 (PRKG2) , P17612 (PRKACA)	Uniprot
S1179	Phosphorylation	P31749 (AKT1)	Uniprot

Research Backgrounds

P29474_NOS3_HUMAN

Function:

Produces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets.

Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.

PTMs:

Phosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity (By similarity). Phosphorylation of Ser-114 by CDK5 reduces activity.

Subcellular Location:

Cell membrane. Membrane>Caveola. Cytoplasm>Cytoskeleton. Golgi apparatus.
Note: Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle; which is favored by interaction with NOSIP and results in a reduced enzymatic activity.

Tissue Specificity:

Platelets, placenta, liver and kidney.

Subunit Structure:

Homodimer. Interacts with NOSIP and NOSTRIN. Interacts with HSP90AB1.

Family&Domains:

Belongs to the NOS family.

Research Fields

· Environmental Information Processing > Signal transduction > Calcium signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > Apelin signaling pathway. (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Metabolism > Amino acid metabolism > Arginine biosynthesis.

· Metabolism > Amino acid metabolism > Arginine and proline metabolism.

· Metabolism > Global and overview maps > Metabolic pathways.

· Organismal Systems > Immune system > Platelet activation. (View pathway)

· Organismal Systems > Endocrine system > Estrogen signaling pathway. (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

References

1). A network pharmacology approach to investigate the mechanism of Shuxuening injection in the treatment of ischemic stroke. Journal of Ethnopharmacology (PubMed: 32315738) [IF=5.4]

Application: WB Species: rat Sample: hippocampal

Fig. 13. |Expression of protein of three genes detected through Western blot. Note: Sham vs I/R, *P < 0.05; SXNI vs I/R, #P < 0.05.

2). Systematic analysis of the mechanism of hydroxysafflor yellow A for treating ischemic stroke based on network pharmacology technology. European Journal of Pharmacology (PubMed: 34302817) [IF=5.0]

Application: IHC Species: Rat Sample: brain tissue

Fig. 12. Immunohistochemical staining and statistical results. The expression levels of positive cells in the hippocampal CA1 area of rats were observed under an optical microscope at a magnification of × 200. Sham versus I/R, # P < 0.05, ## P < 0.01; HSYA versus I/R, **P < 0.01.

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NOS3 Antibody - #DF6450